Biology subjects

Robertson, S. A.

Publications and source records attributed to Robertson, S. A..

3 recordsLinked to original sources

bioRxiv2025

A surface-engineered microfluidic device for antibody-mediated negative selection of high-quality sperm for assisted reproduction

Preparation of spermatozoa with optimal developmental competence remains a challenge in assisted reproduction. Conventional techniques based on sperm motility and morphology fail to adequately remove sperm with DNA damage. Here, we report development of a microfluidic device with a functionalized surface, inspired by the physiological processes of immune cell-mediated sperm selection in the female reproductive tract. A plasma-polymerized polyoxazoline (PPOx) film is applied to glass channel slides by deposition of 2-methyl-2-oxazoline, to establish a stable, biocompatible interface confirmed by X-ray Photoelectron Spectroscopy (XPS), ellipsometry, and sperm culture assays. To selectively eliminate pre-apoptotic and apoptotic spermatozoa wherein DNA damage is common, anti-phosphatidylserine (Anti-PS) antibody is immobilized to the PPOx-coated surface proximal to the channel slide inlet, while the sperm chemoattractant progesterone is adsorbed near the outlet. To optimise selective functionality, the surface topography is tailored by covalent immobilization of gold nanoparticles and addition of microchannels. Sperm recovered after processing whole liquified semen then consistently exhibit high motility and morphology, with <1% showing apoptosis-associated membrane damage or DNA fragmentation. Compared with conventional swim-up or other microfluidic approaches, the device yields sperm with improved quality, offering a simple one-step sperm selection strategy with potential for application in human and animal assisted reproduction. Short text and graphic for 45 the Table of Contents (ToC)This study reports a microfluidic device with a functionalized surface utilizing a polyoxazoline coating and covalently immobilized gold nanoparticles and anti-phosphatidylserine antibody. The device selectively eliminates pre-apoptotic and apoptotic spermatozoa and yields sperm with substantially improved quality and low DNA damage, offering a simple one-step sperm selection device with potential for application in human and animal assisted reproduction. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=166 SRC="FIGDIR/small/673619v1_ufig1.gif" ALT="Figure 1"> View larger version (57K): org.highwire.dtl.DTLVardef@b07026org.highwire.dtl.DTLVardef@192e5b5org.highwire.dtl.DTLVardef@127095aorg.highwire.dtl.DTLVardef@1d70ceb_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

bioRxiv2022

Constitutive expression and distinct properties of IFN-epsilon protect the female reproductive tract from Zika virus infection

The immunological surveillance factors controlling vulnerability of the female reproductive tract (FRT) to sexually transmitted viral infections are not well understood. Interferon-epsilon (IFN{varepsilon}) is a distinct, immunoregulatory type-I IFN that is constitutively expressed by FRT epithelium and is not induced by pathogens like other antiviral IFNs , {beta} and {lambda}. We show the necessity of IFN{varepsilon} for Zika Virus (ZIKV) protection by: increased susceptibility of IFN{varepsilon}-/- mice; their "rescue" by intravaginal recombinant IFN{varepsilon} treatment and blockade of protective endogenous IFN{varepsilon} by neutralising antibody. Complementary studies in human FRT cell lines showed IFN{varepsilon} had potent anti-ZIKV activity, associated with transcriptome responses similar to IFN{lambda} but lacking the proinflammatory gene signature of IFN. IFN{varepsilon} activated STAT1/2 pathways similar to IFN and {lambda} that were inhibited by ZIKV-encoded non-structural (NS) proteins, but not if IFN{varepsilon} exposure preceded infection. This scenario is provided by the constitutive expression of endogenous IFN{varepsilon}. However, the IFN{varepsilon} expression was not inhibited by ZIKV NS proteins despite their ability to antagonise the expression of IFN{beta} or {lambda}. Thus, the constitutive expression of IFN{varepsilon} provides cellular resistance to viral strategies of antagonism and maximises the antiviral activity of the FRT. These results show that the unique spatiotemporal properties of IFN{varepsilon} provides an innate immune surveillance network in the FRT that is a significant barrier to viral infection with important implications for prevention and therapy. Author SummaryThe female reproductive tract (FRT) is vulnerable to sexually transmitted infections and therefore a well-tuned immune surveillance system is crucial for maintaining a healthy FRT. However, our understanding of the factors that impact viral infection of the FRT and the host response are not well understood. In this work we investigate the role of a hormonally regulated type I interferon, IFN epsilon (IFN{varepsilon}) in control of Zika virus (ZIKV) infection of the FRT. IFN{varepsilon} is unique compared to other canonical type-I IFNs in that it is constitutively expressed by epithelial cells of the FRT with expression levels controlled by progesterone and not in response to viral infection. We demonstrate that IFN{varepsilon} has anti-ZIKV properties using a combination of IFN{varepsilon} KO mice, blockade of endogenous IFN{varepsilon} by neutralising Abs and rescue of IFN{varepsilon} KO mice by recombinant IFN{varepsilon} administered directly to the FRT. Furthermore, we complemented our in vivo studies using human FRT derived cell lines. Importantly, ZIKV NS proteins did not block IFN{varepsilon} expression despite their ability to antagonise the expression of IFN{beta} or {lambda}. Collectively this work implicates IFN{varepsilon} as a key type-I IFN that provides a distinct homeostatic antiviral environment in the FRT.

immunology↗

bioRxiv2017

Transplacental protection against asthma by maternal treatment with a bacterial-derived immunomodulatory agent

Studies in European and US farming populations have documented major reductions in asthma prevalence in offspring of mothers exposed to microbial breakdown products present in farm dusts and unprocessed foods. This was associated with enhancement of innate immune competence in the offspring. We sought to (i) identify a safe therapeutic that would reproduce these immunomodulatory effects in a murine model, (ii) elucidate underlying mechanism(s)-of-action, and (iii) develop a scientific rationale for progressing this approach to human trials. We demonstrate in mice that maternal treatment during pregnancy with the microbial-derived immunomodulator OM85, which has been used clinically in adults and children in Europe for >30 years for bolstering resistance to infection-associated airways inflammation, markedly reduces the susceptibility of the offspring of treated mothers to development of experimental atopic asthma. We identify bone marrow precursors of the dendritic cell populations responsible for airway mucosal immune surveillance as the primary targets for the asthma-protective effects of maternal OM85 treatment in the offspring.

immunology↗